Ca-deficient hydroxyapatite/polylactide nanocomposites with chemically modified interfaces by high pressure consolidation at room temperature

Hydroxyapatite (HAP) is a close synthetic analog of the bone mineral and is often considered as a material for bone graft substitutes and tissue engineering scaffolds. Despite its attractive bioactive properties low-fracture toughness limits the use of HAP ceramics to a number of non-load-bearing applications. To obtain a more adequate mechanical behavior, HAP is often combined with polymers based on lactic and glycolic acids or polycaprolactone using hot pressing. In such composite materials, the compatibility and bonding strength of HAP–polymer interfaces are critical parameters that must be controlled and improved. This may be achieved, for example, by covalent immobilization of organic moieties on the ceramic particles surface. In this work, the surface of calcium-deficient hydroxyapatite (CDHAP) was modified by reaction with hexamethylene diisocyanate (HDI) in a non-aqueous suspension. Composites of CDHAP–HDI with polylactide (PLA) were high pressure consolidated at room temperature at 2.5 GPa yielding up to 90% theoretical density. The effects of total organic fraction and modification extent on compression strength were studied. Materials with high extent of modification and high organic content exhibited compressive strength of ~295 MPa, much higher than reported in other studies. These materials are suitable candidates for load bearing orthopedic applications.     

An <Emphasis Type="Italic">in vivo</Emphasis> study of a bone grafting material consisting of hydroxyapatite and reconstituted collagen

This study aims to evaluate the performance of our recently developed microspheres of hydroxyapatite/reconstituted collagen as a bone grafting material. The microspheres were fabricated into a circular disc and implanted in a pre-drilled hole in a rats calvaria. The bone tissue had regenerated and grown into the disc bone graft 4 weeks following implantation. After 16 weeks of implantation, the regenerated bone had integrated with the remaining material and made close contact with it. The disc had been completely absorbed with almost no visible bone graft left after 24 weeks of implantation. In contrast, a hydroxyapatite disc still remained intact on the 24th week after implantation. These results suggested that the hydroxyapatite/reconstituted collagen microsphere can be used as an excellent bone grafting material.     

In vivo evaluation of bone tissue behavior on ion implanted surfaces

The ion implantation process offers several unique advantages over other surfaces modifications techniques, in regard to ion release and material mechanical characteristics. The aim of this study was to evaluate the in vivo bone tissue response to ion implanted surfaces. Untreated and nitrogen-ion-implanted stainless steel implants were inserted in the tibia diaphysis (cortical bone) and proximal tibia epiphysis (trabecular bone) of 12 New Zealand White rabbits. The animals were divided into three groups of four animals each, which were maintained for 4, 12 and 24 weeks according to internationally accepted and standardized procedures. At sacrifice, the implants were retrieved with surrounding bone and fixed in 4% neutral buffered formaldehyde and embedded in polymethylmethacrylate (PMMA). The samples were reducted in slices and stained with hematoxylin-eosin, light-green, fuchsin acid and giemsa solution for histological evaluation; fluorescent markers were also used to assess bone apposition. Histomorphometric evaluation was used to determine the extent of bone-material contact. Results from histological and morphometrical analyses revealed active remodeling of bone around both types of implants (control and ion implanted). However, faster bone deposition was observed around the treated material (12 weeks). Both materials reached similar endpoints, as no significant differences between them were evident at 24 weeks. The results demonstrate that ion implanted stainless steel has similar, or slightly enhanced, biological compatibility in contact with bone compared to untreated material; thus it may be a useful material in biomedical applications where reduced ion release or enhanced mechanical properties (as provided by ion implantation) are required. © 2001 Kluwer Academic Publishers     

Preparation of electrospun siloxane-poly(lactic acid)-vaterite hybrid fibrous membranes for guided bone regeneration

A nonwoven fabric scaffold for guided bone regeneration (GBR) consisting of siloxane-poly(lactic acid) (PLA)-vaterite hybrid material (SiPVH) was prepared by an electrospinning method. To improve the cellular compatibility of the fabric, the fibers were completely coated with hydroxyapatite (HA) by soaking in simulated body fluid. The HA-coated SiPVH nonwoven fabric contained large-sized spaces and showed the ability of releasing soluble silica and calcium species, which have previously been reported to stimulate osteogenic cells at the genetic level. A new type of GBR membrane was prepared by bonding SiPVH fabric with PLA nonwoven fabric with small-sized pores for preventing the intrusion of soft tissue. The resultant bi-layered membrane was expected to be effective not only for having an open structure for bone formation and a barrier to soft tissue, but also for enhancing bone growth by the release of ionic species.     

Novel porous scaffolds of poly(lactic acid) produced by phase-separation using room temperature ionic liquid and the assessments of biocompatibility

Here we prepared three-dimensional (3D) porous-structured biodegradable polymer scaffolds for tissue regeneration using room temperature ionic liquids (RTILs) as a novel porogen, and addressed their biological properties, including in vitro cell growth and differentiation and in vivo tissue compatibility. RTIL based on 1-butyl-3-methylimidazolium ([bmim]) bearing hydrophilic anion Cl was introduced within the polymer structure to provide a pore network. A mixture of poly(lactic acid) (PLA) with RTIL dissolved in an organic solvent formed a bi-continuous network during the drying process. Selective dissolution of the RTIL phase was facilitated in ethanol, which resulted in a porous network of the polymer phase with complete removal of the RTIL. The RTILs-assisted porous scaffolds showed a typical open-channeled network with pore sizes over 100 μm and porosities of about 86–94%. For the biocompatibility assessments of the scaffolds, mesenchymal stem cells (MSCs) derived from rat bone marrow were seeded onto the PLA scaffold, and the cell proliferation and osteoblastic differentiation behaviors were examined. Results showed a typical on-going increase in the cell population with a level comparable to that observed on the tissue culture plastic control, indicating good cell compatibility. When cultured in an osteogenic medium, the alkaline phosphatase (ALP) activity of the cells on the PLA scaffolds was stimulated to increase with time from 7 to 14 days, in a similar manner to that on the control. Moreover, the expression of genes related to osteoblasts, including collagen type I, osteocalcin and bone sialoprotein, was stimulated on the 3D PLA scaffold during culture for up to 14 days, with levels higher than those on the control, suggesting the developed scaffold provided a 3D matrix condition for osteogenesis. An in vivo pilot study conducted subcutaneously in rat for 4 weeks revealed good tissue compatibility of the scaffold, with the ingrowth of cells and formation of collageneous tissue around and deep within the pores of the scaffold and no significant inflammatory reaction. Taken together, this novel method of using RTILs as a pore generator is considered to be useful in the development of biocompatible porous polymer scaffolds for tissue regeneration.     

Gamma-irradiated EPR response of nano-structure hydroxyapatite synthesised via hydrolysis method

In this paper, the gamma-irradiated EPR responses of hydroxyapatite (HAP) samples were investigated from a dosimetric point of view. For this purpose, nano-structure hydroxyapatite was synthesised via a hydrolysis method. A portion of the produced powder was annealed at 600 °C for 4 h. All the samples were irradiated under the (60)Co gamma-ray source at different absorbed doses from 0.1 to 45 kGy. EPR signal intensities of HAP samples were measured at room temperature in air. Subsequently, the EPR signal intensities were constructed as a function of radiation dose and were compared with the results of bone powder samples. The results show that the EPR responses of non-annealed HAP samples are higher than others for several times.     

Tissue reactions to bioabsorbable ciprofloxacin-releasing polylactide-polyglycolide 80/20 screws in rabbits’ cranial bone

The aim of this study was to assess tissue reactions to bioabsorbable self-reinforced ciprofloxacin-releasing polylactide/polyglycolide (SR-PLGA) 80/20 screws in rabbits' cranial bone. Two screws were implanted in each rabbit, one screw on either side of the sagittal suture (n = 28 rabbits). Animals were sacrificed after 2, 4, 8, 16, 24, 54 and 72 weeks, four animals per group. On histological examination the number of macrophages, giant cells, active osteoblasts and fibrous tissue layers were assessed and degradation of the screws was evaluated. At 2 weeks, the highest number of macrophages and giant cells were seen near the heads of the screws. After 4 and 8 weeks, the number of giant cells decreased but that of macrophages decreased from 16 weeks and on. Screws were surrounded by fibrous tissue capsule that progressively was growing in thickness by time. Active osteoblasts were seen around the shaft of the screws with the highest number seen at 4 weeks postoperatively. At 16 weeks, compact fragmentation of the screw heads was seen with macrophages seen inside the screw matrices. After 24 weeks, no polarization of the screws was seen. After one year, PLGA screws had been replaced by adipose tissue, fibrous tissue and "foamy macrophages" which had PLGA particles inside them. After 1(1/2) years, the amount of biomaterial remaining had decreased remarkably. The particles of biomaterial were inside "foamy macrophages." Ciprofloxacin-releasing SR-PLGA 80/20 screws elicited a mild inflammatory reaction but did not interfere with osteoblast activity. No complications were seen when implanted in cranial bone of rabbit.     

Biological response of new activated acrylic bone cements with antiseptic properties. Histomorphometric analysis

The biological response to an acrylic bone cement cured with 4,4′-bis-dimethylamino benzydrol (BZN) as activator of reduced cytotoxicity and antiseptic properties, has been carried out and compared with that obtained for CMW 3 cement. Histomorphometrical data (undecalcified trichromic Goldner staining) were obtained by measuring the most significant variables at the bone-cement interface. Quantitative results of tissue response revealed that newly formed bone and connective tissue were maximum at 4 weeks whereas bone marrow increased with time of implantation for both cements. Statistical analysis (p < 0.05) showed no significant differences in newly formed bone and bone marrow with time and between both groups, however, connective tissue significantly decreased between 4 weeks and 12 weeks for BZN cement, and between 12 weeks and 24 weeks for CMW3. By comparing both cements at each time, lower significant percentage of connective tissue at the bone-cement interface of the BZN cement, was obtained at 12 and 24 weeks, however, a very low amount of connective tissue was found for both cements. All the results indicate that the new activated system could be applied clinically in a relatively short time, after the corresponding preclinical study.     

原位沉析羟基磷灰石-壳聚糖骨组织工程支架材料的研制

仿生构建羟基磷灰石-壳聚糖复合材料是制备骨组织修复材料的重要途径之一.本研究利用有机官能团对无机物矿化的调控作用,在壳聚糖多孔支架表面原位沉析羟基磷灰石(HAp).在仿生溶液中,采用简单的化学处理,使HAp晶体在壳聚糖多孔支架表面原位沉析.研究结果表明壳聚糖分子结构中的氨基作为成核位点,在碱性条件下首先与吸附Ca2 ,再通过静电作用力吸附仿生溶液中的PO43-、OH-等其它离子促使HAp晶体在壳聚糖支架材料表面的成核、长大.此类材料有望成为一种生物活性的骨组织工程材料.     

Tissue responses to anti-washout apatite cement using chitosan when implanted in the rat tibia

The tissue response to anti-washout apatite cement using chitosan (aw-AC(chi)) was evaluated by implanting aw-AC(chi) into bone defects of rat tibiae using conventional apatite cement (c-AC) as a control material. During the experimental period up to 16 weeks, the only difference between aw-AC and c-AC was found at two weeks in the tissue response of soft tissue. At two weeks, c-AC showed a moderate inflammatory response; small particles of c-AC were scattered in the cutaneous tissue and many foreign body giant cells were collected around the scattered c-AC, whereas aw-AC showed only a slight inflammatory response and few foreign body giant cells. We found no difference between aw-AC(chi) and c-AC with respect to bone tissue response. Both AC were almost completely surrounded by mature bone at eight weeks. No promotion or reduction of osteoconductivity was observed by chitosan even though it is considered to promote bone formation.We concluded, therefore, that enhancement of bone formation cannot be expected by employing chitosan to obtain anti-washout properties, at least in the concentration used in this study, even though aw-AC(chi) is much more useful than c-AC. © 2001 Kluwer Academic Publishers     

纳米羟基磷灰石/聚酰胺6医用复合材料的制备及性能表征

为防止纳米羟基磷灰石(nano HAP)粉末的团聚,采用溶剂沉淀法制备了nano HAP/聚酰胺6(PA6)复合粉末,并对粉末进行热压成型制得nano HAP/PA6复合材料。然后,通过FTIR、XRD和SEM对nano HAP/PA6复合材料的成分、结构和形貌进行了表征,并对复合材料的热稳定性、力学性能和细胞相容性进行了检测。结果表明:所制备的nano HAP/PA6复合材料结晶体大小均匀,且PA6只存在α型结晶;由于nano HAP与PA6界面上形成新的氢键和COO—Ca,复合材料具有良好的综合性能;在低于350℃时,nano HAP/PA6复合材料不会发生裂解,力学性能与人骨匹配,50wt%nano HAP/PA6复合材料的弯曲强度、压缩强度和弹性模量分别为146.87MPa、98.44MPa和5.44GPa。MG-63骨瘤细胞在nano HAP/PA6复合材料表面粘附和生长状况良好,说明nano HAP/PA6复合材料具有良好的细胞相容性。所得结论表明nano HAP/PA6复合材料在骨修复方面具有应用价值。     

A novel, low temperature method for the preparation of ß-TCP/HAP biphasic nanostructured ceramic scaffold from natural cancellous bone

In this study, a novel method was used to synthesize ß-TCP/HAP biphasic ceramic scaffold from natural cancellous bone. Bovine bone was calcined to remove the organic content. The remaining material was then soaked in P₂O₅ solution for different periods. P ions were doped into cancellous bone and reacted with hydroxyapatite (HAP) to produce ß-tricalcium phosphate (ß-TCP) at room temperature without any subsequent high-temperature heat treatment. By increasing the soaking time, the crystalline phase composition of the calcined bone gradually changed from HAP into a HAP/ß-TCP biphasic structure with different HAP/TCP ratios. The crystallite size of HAP and ß-TCP was smaller than 100 nm.     

Porous polymer/hydroxyapatite scaffolds: characterization and biocompatibility investigations

Poly-lactic-glycolic acid (PLGA) has been widely used as a scaffold material for bone tissue engineering applications. 3D sponge-like porous scaffolds have previously been generated through a solvent casting and salt leaching technique. In this study, polymer–ceramic composite scaffolds were created by immersing PLGA scaffolds in simulated body fluid, leading to the formation of a hydroxyapatite (HAP) coating. The presence of a HAP layer was confirmed using scanning electron microscopy, energy dispersive X-ray spectroscopy and Fourier transform infrared spectroscopy in attenuated total reflection mode. HAP-coated PLGA scaffolds were tested for their biocompatibility in vitro using human osteoblast cell cultures. Biocompatibility was assessed by standard tests for cell proliferation (MTT, WST), as well as fluorescence microscopy after standard cell vitality staining procedures. It was shown that PLGA–HAP composites support osteoblast growth and vitality, paving the way for applications as bone tissue engineering scaffolds.     

HAP／Y－TZP复相生物陶瓷研究

羟基磷灰石，是一种具有优秀生物活性的生物材料，但作为承受负荷的骨组织替代物时，缺乏良好的力学性能．本文采用湿化学法制备ＨＡＰ及ＨＡＰ／Ｙ－ＴＺＰ复相粉体，通过热压工艺，以期改善ＨＡＰ的力学性能，分别进行上皮组织、成纤维细胞与膜片联合培养试验．结果表明：含４０ｗｔ％Ｙ－ＴＺＰ的ＨＡＰ复相陶瓷，其抗弯强度和断裂韧性均比同种工艺得到的ＨＡＰ提高７４％和９０％；上皮组织与膜片贴覆性好，不延缓细胞的生长，成纤维细胞与膜片接种良好．     

Bonding ability evaluation of bone cement on the cortical surface of rabbit’s tibia

A composite bone cement designated G2B1 that contains β tricalcium phosphate particles was developed as a bone substitute for percutaneous transpedicular vertebroplasty. In this study, both G2B1 and commercial PMMA bone cement (CMW1) were implanted into proximal tibiae of rabbits, and their bone-bonding strengths were evaluated at 4, 8, 12 and 16 weeks after implantation. Some of the specimens were evaluated histologically using Giemsa surface staining, contact microradiography (CMR) and scanning electron microscopy (SEM). Histological findings showed that G2B1 contacted bone directly without intervening soft tissue in the specimens at each time point, while there was always a soft tissue layer between CMW1 and bone. The bone-bonding strength of G2B1 was significantly higher than that of CMW1 at each time point, and significantly increased from 4 weeks to 8 and 12 weeks, while it decreased significantly from 12 weeks to 16 weeks. Bone remodeling of the cortex under the cement was observed especially for G2B1 and presumably influenced the bone bonding strength of the cement. The results indicate that G2B1 has bioactivity, and bone bonding strength of bioactive bone cements can be estimated fairly with this experimental model in the short term.     

Preliminary<Emphasis Type="Italic">in vitro</Emphasis> and<Emphasis Type="Italic">in vivo</Emphasis> characterizations of a sol-gel derived bioactive glass-ceramic system

This study investigates quantitatively and qualitatively the sol-gel derived bioactive glass-ceramic system (BGS)—apatite-wollastonite (AW) type granules in the size range of 0.5–1 mm, as an effective graft material for bone augmentation and restoration. Scanning electron micrographs (SEM) of the sintered granules revealed the rough material surface with micropores in the range 10–30 μm. X-ray diffraction (XRD) pattern of the granules revealed the presence of crystalline phases of the hydroxyapatite and wollastonite, and the functional groups of the silicate and phosphates were identified by Fourier transform infrared spectroscopy (FT-IR). Thein vitro cell culture studies with L929 mouse fibroblast cell line showed very few cells adhered on the BGS disc after 24 h. This could be due to the highly reactive surface of the disc concomitant with the crystallization but not due to the cytotoxicity of the material, since the cellular viability (MTT assay) with the material was 80‰ Cytotoxicity and cytocompatibility studies proved that the material was non-toxic and biocompatible. After 12 weeks of implantation of the BGS granules in the tibia bone of New Zealand white rabbits, the granules were found to be well osteointegrated, as observed in the radiographs. Angiogram with barium sulphate and Indian ink after 12 weeks showed the presence of microcapillaries in the vicinity of the implant site implicating high vascularity. Gross observation of the implant site did not show any inflammation or necrosis. SEM of the implanted site after 24 weeks revealed good osteointegration of the material with the newly formed bone and host bone. New bone was also observed within the material, which was degrading. Histological evaluation of the bone healing with the BGS granules in the tibial defect at all time intervals was without inflammation or fibrous tissue encapsulation. After 2 weeks the new bone was observed as a trabeculae network around the granules, and by 6 weeks the defect was completely closed with immature woven bone. By 12 weeks mature woven bone was observed, and new immature woven bone was seen within the cracks of the granules. After 24 weeks the defect was completely healed with lamellar bone and the size of the granules decreased. Histomorphometrically the area percentage of new bone formed was 67.77% after 12 weeks and 63.37% after 24 weeks. Less bone formation after 24 weeks was due to an increased implant surface area contributed by the material degradation and active bone remodeling. The osteostimulative and osteoconductive potential of the BGS granules was established by tetracycline labelling of the mineralizing areas by 2 and 6 weeks. This sol-gel derived BGS granules proved to be bioactive and resorbable which in turn encouraged active bone formation.     

Novel osteoconductive β-tricalcium phosphate/poly(L-lactide-co-e-caprolactone) scaffold for bone regeneration: a study in a rabbit calvarial defect

The advantages of synthetic bone graft substitutes over autogenous bone grafts include abundant graft volume, lack of complications related to the graft harvesting, and shorter operation and recovery times for the patient. We studied a new synthetic supercritical CO₂ –processed porous composite scaffold of β-tricalcium phosphate and poly(L-lactide-co-caprolactone) copolymer as a bone graft substitute in a rabbit calvarial defect. Bilateral 12?mm diameter critical size calvarial defects were successfully created in 18 rabbits. The right defect was filled with a scaffold moistened with bone marrow aspirate, and the other was an empty control. The material was assessed for applicability during surgery. The follow-up times were 4, 12, and 24 weeks. Radiographic and micro-CT studies and histopathological analysis were used to evaluate new bone formation, tissue ingrowth, and biocompatibility. The scaffold was easy to shape and handle during the surgery, and the bone-scaffold contact was tight when visually evaluated after the implantation. The material showed good biocompatibility and its porosity enabled rapid invasion of vasculature and full thickness mesenchymal tissue ingrowth already at four weeks. By 24 weeks, full thickness bone ingrowth within the scaffold and along the dura was generally seen. In contrast, the empty defect had only a thin layer of new bone at 24 weeks. The radiodensity of the material was similar to the density of the intact bone. In conclusion, the new porous scaffold material, composed of microgranular β-TCP bound into the polymer matrix, proved to be a promising osteoconductive bone graft substitute with excellent handling properties.

Open image in new window

     

Tissue response to hafnium

The aim of the present experimental study was to evaluate the tissue response to hafnium (Hf) a reactive metal closely related to titanium (Ti) and zirconium (Zr). Hf has not been previously evaluated as implant material in a biologic environment. In a first experiment, 21 machined Hf non-threaded implants (test) and 21 similar Ti implants (control) were inserted in the abdominal wall of 21 rats. Animals were sacrificed after 8 days (6 rats), 6 (7 rats) and 12 weeks (8 rats). In a second experiment, 18 rabbits received 18 Hf and 18 Ti threaded implants in their tibiae, one implant in each tibia. The rabbits were sacrificed after 6, 12 and 24 weeks (6 animals/time interval). The bulk metal of the abdominal wall implants, embedded together with the surrounding tissue, was electrolytically dissolved and semithin (1 m) sections of the intact tissue–implant interface were evaluated by light microscopy (morphometry). Bone-implant contact and bone area within threads were evaluated in ground sections. In soft tissues, a fluid space containing predominantly monocytes/macrophages surrounded the abdominal implants at 8 days. At 6 and 12 weeks, a fibrous capsule, consisting of layers of macrophages and fibroblasts, surrounded the implants. Macrophages, including multinuclear giant cells, always formed the innermost layer in contact with the implant surface. No quantitative or qualitative difference in the tissue organization was detected between Ti and Hf implants. In rabbits, 6 weeks after insertion, the proximal two threads located within the cortical bone were filled with bone in contact with Hf and Ti. The distal threads contained bone marrow. After 12 and 24 weeks, mature bone was present in the proximal 3–4 implant threads. No statistically significant difference was found between Hf and Ti implants at any time periods. It is concluded that Hf is an interesting metal for biomedical applications in bone and soft tissue. © 2001 Kluwer Academic Publishers     

Tissue response to a braided poly-L-lactide implant in an experimental reconstruction of anterior cruciate ligament

During follow-up periods of 6, 12, 24 and 48 weeks, the tissue response to a braided poly-L-lactide (PLLA) implant, 3.2 mm in diameter, was investigated in the reconstruction of experimental anterior cruciate ligament (ACL) ruptures in 32 sheep. In 16 sheep the cut ACL was removed and reconstructed with the fascia lata augmented with a PLLA implant. In 16 sheep the ACL was cut from its midportion, sutured, and thereafter augmented with a PLLA implant. The tissue reactions were typical of a scant non-specific-foreign-body reaction. The number of inflammatory and giant cells was greatest at six weeks, decreasing thereafter. Degradation of the PLLA was incomplete at 48 weeks. No signs of synovitis or changes in the cartilaginous surfaces were observed. The reconstructions in both groups were anchored to the bone by fibroconnective tissue, and remodelling of the bone was seen along the drill channels. After 48 weeks the maturation of the fibroconnective tissue and the orientation of the collagen fibres were higher (p<0.01) in the fascia-lata-PLLA group than in the primarysuture-PLLA group. Histologically, the braided PLLA implants proved to be suitable for ACL repair in sheep. The augmentation of the fascia lata with the PLLA implant seemed to be preferable to that of the primary suture of the ACL.     

Determination of relative in vivo osteoconductivity of modified potassium fluorrichterite glass–ceramics compared with 45S5 bioglass

Potassium fluorrichterite (KNaCaMg₅Si₈O₂₂F₂) glass–ceramics were modified by either increasing the concentration of calcium (GC5) or by the addition of P₂O₅ (GP2). Rods (2?×?4?mm) of stoichiometric fluorrichterite (GST), modified compositions (GC5 and GP2) and 45S5 bioglass, which was used as the reference material, were prepared using a conventional lost-wax technique. Osteoconductivity was investigated by implantation into healing defects in the midshaft of rabbit femora. Specimens were harvested at 4 and 12?weeks following implantation and tissue response was investigated using computed microtomography (μCT) and histological analyses. The results showed greatest bone to implant contact in the 45S5 bioglass reference material at 4 and 12?weeks following implantation, however, GST, GC5 and GP2 all showed direct bone tissue contact with evidence of new bone formation and cell proliferation along the implant surface into the medullary space. There was no evidence of bone necrosis or fibrous tissue encapsulation around the test specimens. Of the modified potassium fluorrichterite compositions, GP2 showed the greatest promise as a bone substitute material due to its osteoconductive potential and superior mechanical properties.